Deuterated amlexanox

ABSTRACT

Provided herein is technology relating to deuterated amlexanox and particularly, but not exclusively, to compositions comprising deuterated amlexanox, methods of producing deuterated amlexanox, and uses of deuterated amlexanox.

This application is a continuation of U.S. application. Ser. No.15/153,405, filed May 12, 2016, which is a divisional application ofU.S. application. Ser. No. 14/888,591, filed Nov. 2, 2015, now U.S. Pat.No. 9,365,581, Issued Jun. 14, 2016, which is a 371 U.S. National PhaseEntry of International Application No. PCT/US2014/036644, filed May 2,2014, which claims priority to U.S. Pat. Appl. Ser. No. 61/818,753,filed May 2, 2013, each of which is incorporated herein by reference inits entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under DK060591 awardedby the National Institutes of Health. The government has certain rightsin the invention.

FIELD OF INVENTION

Provided herein is technology relating to deuterated amlexanox andparticularly, but not exclusively, to compositions comprising deuteratedamlexanox, methods of producing deuterated amlexanox, and uses ofdeuterated amlexanox.

BACKGROUND

The incidence of the metabolic disorders of diabetes and obesity hasreached epidemic levels. It has been estimated that over 120 millionAmericans are clinically over-weight and over 25 million have diabetes,including 1.9 million new cases in 2010 among those aged 20 and older.Obesity and diabetes can cause or contribute to the development of, orat least affect the treatment of, other diseases and disorders such ascardiovascular diseases, stroke, hypertension, kidney failure, asthma,and cancer. The economic burden of diabetes alone was estimated to beover $174 billion per year in 2007. Obesity and diabetes have a majorimpact on human health and the various national healthcare systems allover the world.

Recently launched weight-loss drugs have failed or have demonstratedlimited efficacy and undesirable side effects. Similarly, despite atremendous medical need, the pharmaceutical industry has realized onlylimited success developing therapeutics to manage diabetes. The mostcommon therapeutics (sulfonylureas) are not effective and the mostpromising new drugs (thiazolidinediones) have demonstrated rare butfatal side effects. Thus, there is an urgent need for a morecomprehensive understanding of the molecular basis of obesity anddiabetes, for tests that allow early detection of predispositions to thedisorders, and for more effective pharmaceuticals for preventing andtreating these diseases and conditions.

SUMMARY

Accordingly, provided herein are deuterated amlexanox compounds for thetreatment of obesity, insulin resistance, diabetes, and steatosis. Inaddition, the deuterated compounds are anti-inflammatory antiallergicimmunomodulators, e.g., for the treatment of diseases associated withinflammation. The deuterium kinetic isotope effect associated withplacing deuterium at the site of metabolic derivatization slowsmetabolic derivatization and thus increases the lifetime of the activedrug in vivo. The deuterated amlexanox derivatives provided herein weredemonstrated to have biological activity in inhibiting protein kinases(e.g., TBK1 or IKKε) associated with disease.

Accordingly, provided herein is technology related to a compositioncomprising deuterated amlexanox or a pharmaceutically acceptable saltthereof. In some embodiments, the composition comprising deuteratedamlexanox or a pharmaceutically acceptable salt thereof furthercomprises non-deuterated amlexanox or a pharmaceutically acceptable saltthereof. Some embodiments provide a composition comprising a compoundhaving a structure according to:

or a pharmaceutically acceptable salt thereof, wherein X, Y, or Z is agroup that comprises deuterium (D). In some embodiments, X is a groupthat comprises D; in some embodiments, X and Y are groups that compriseD; and in some embodiments X, Y, and Z are groups that comprise D. Insome embodiments, X, Y, or Z comprises or is CD₃; in some embodiments,X, Y, or Z comprises or is D; in some embodiments, X, Y, or Z comprisesor is CH₂CDH₂; in some embodiments, X and Y comprise or are CD₃ and Zcomprises or is D; in some embodiments, X and Y comprise or are CH₃ andZ comprises or is D; and in some embodiments, X comprises or is CH₃, Ycomprises or is CH₂D, and Z comprises or is D.

Some embodiments provide a compound having the structural formula:

or a pharmaceutically acceptable salt thereof, wherein X, Y, or Z isenriched with deuterium more than 10%. In some embodiments, X, Y, or Zis enriched with deuterium more than 20%, 30%, 40%, 50%, 60%, 80%, 90%,95%, 98%, or 99% or more.

In some embodiments are provided a composition comprising a compound orcomposition as described herein and a pharmaceutically acceptablecarrier.

In some embodiments, the technology is related to use of a compositioncomprising a compound having a structure according to:

or a pharmaceutically acceptable salt thereof, for a medicament, whereinX, Y, or Z is a group that comprises D. Furthermore, in someembodiments, the technology provides use of a composition as providedherein and/or use of a compound as provided herein. Some embodimentsprovide use of a composition comprising a compound having a structureaccording to:

for a medicament to treat diabetes, insulin resistance, steatosis,hepatitis, obesity, allergic rhinitis, conjunctivitis, allergy, asthma,immune disorder, atherosclerosis, canker sore, ulcer, aphthous ulcer,symptoms of Behçet's Disease, or inflammation, wherein X, Y, or Z is agroup that comprises D.

Some embodiments provide a method of treating a subject comprisingadministering to the subject a deuterated amlexanox. Some embodimentsprovide a method of treating a subject comprising administering to thesubject a composition comprising a compound having a structure accordingto:

or a pharmaceutically acceptable salt thereof, wherein X, Y, or Z is agroup that comprises D.

In some embodiments, the technology provides a method of treating amalady that is diabetes, insulin resistance, steatosis, hepatitis,obesity, allergic rhinitis, conjunctivitis, allergy, asthma, immunedisorder, atherosclerosis, canker sore, ulcer, aphthous ulcer, symptomsof Behçet's Disease, or inflammation comprising identifying a subject inneed of a treatment for the malady and administering to the subject acomposition comprising a compound having a structure according to:

or a pharmaceutically acceptable salt thereof, wherein X, Y, or Z is agroup that comprises D. In some embodiments, the technology provides amethod of inhibiting a TBK1 or IKKε kinase in a patient comprisingselecting a patient in need of a compound to inhibit a TBK1 or IKKεkinase and administering to the subject a composition comprising acompound having a structure according to:

or a pharmaceutically acceptable salt thereof, wherein X, Y, or Z is agroup that comprises D.

Some embodiments provide a method of manufacturing a deuteratedamlexanox comprising providing a deuterated precursor and synthesizingthe deuterated amlexanox from the deuterated precursor. In someembodiments the deuterated precursor is a deuterated alcohol and in someembodiments the deuterated precursor is R₂CDOH.

Some embodiments provide a deuterated amlexanox produced by providing adeuterated precursor and synthesizing the deuterated amlexanox from thedeuterated precursor. In some embodiments the deuterated precursor is adeuterated alcohol and in some embodiments the deuterated precursor isR₂CDOH. Additional embodiments will be apparent to persons skilled inthe relevant art based on the teachings contained herein.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features, aspects, and advantages of the presenttechnology will become better understood with regard to the followingdrawings:

FIG. 1 is a plot showing a dose-dependent inhibition of IKKε and TBK1 byembodiments of the compounds described herein.

It is to be understood that the figures are not necessarily drawn toscale, nor are the objects in the FIGURES necessarily drawn to scale inrelationship to one another. The figures are depictions that areintended to bring clarity and understanding to various embodiments ofapparatuses, systems, and methods disclosed herein. Wherever possible,the same reference numbers will be used throughout the drawings to referto the same or like parts. Moreover, it should be appreciated that thedrawings are not intended to limit the scope of the present teachings inany way.

DETAILED DESCRIPTION

Provided herein are new amlexanox compounds, pharmaceutical compositionsmade thereof, and methods to treat disease, e.g., by inhibiting kinaseIKKε or TBK1) activity, in a subject. The technology finds use in thetreatment of disorders such as diabetes, insulin resistance, steatosis,hepatitis, obesity, allergic rhinitis, conjunctivitis, allergy, asthma,immune disorder, atherosclerosis, canker sore, ulcer (e.g., aphthousulcer, symptoms of Behçet's Disease, etc.), or inflammation (e.g.,inflammatory bowel disease, Crohn's disease, osteoarthritis, etc.).

In addition, the compounds find use to treat subjects affected withinflammatory diseases or disorders such as gout, arthritis (e.g., acuteor chronic idiopathic inflammatory arthritis, osteoarthritis),psoriasis, chronic dermatosis, myositis, demyelinating diseases, chronicobstructive pulmonary disease (COPD), interstitial lung disease,glomerulonephritis, interstitial nephritis, chronic active hepatitis,Crohn's disease, inflammatory bowel disease, ulcerative colitis, plaqueformation in atherosclerosis, degenerative diseases of the joints ornervous system, or multiple sclerosis (MS).

The section headings used herein are for organizational purposes onlyand are not to be construed as limiting the described subject matter inany way.

In this detailed description of the various embodiments, for purposes ofexplanation, numerous specific details are set forth to provide athorough understanding of the embodiments disclosed. One skilled in theart will appreciate, however, that these various embodiments may bepracticed with or without these specific details. In other instances,structures and devices are shown in block diagram form. Furthermore, oneskilled in the art can readily appreciate that the specific sequences inwhich methods are presented and performed are illustrative and it iscontemplated that the sequences can be varied and still remain withinthe spirit and scope of the various embodiments disclosed herein.

All literature and similar materials cited in this application,including but not limited to, patents, patent applications, articles,books, treatises, and internet web pages are expressly incorporated byreference in their entirety for any purpose. Unless defined otherwise,all technical and scientific terms used herein have the same meaning asis commonly understood by one of ordinary skill in the art to which thevarious embodiments described herein belongs. When definitions of termsin incorporated references appear to differ from the definitionsprovided in the present teachings, the definition provided in thepresent teachings shall control.

Definitions

To facilitate an understanding of the present technology, a number ofterms and phrases are defined below. Additional definitions are setforth throughout the detailed description.

Throughout the specification and claims, the following terms take themeanings explicitly associated herein, unless the context clearlydictates otherwise. The phrase “in one embodiment” as used herein doesnot necessarily refer to the same embodiment, though it may.Furthermore, the phrase “in another embodiment” as used herein does notnecessarily refer to a different embodiment, although it may. Thus, asdescribed below, various embodiments of the invention may be readilycombined, without departing from the scope or spirit of the invention.

In addition, as used herein, the term “or” is an inclusive “or” operatorand is equivalent to the term “and/or” unless the context clearlydictates otherwise. The term “based on” is not exclusive and allows forbeing based on additional factors not described, unless the contextclearly dictates otherwise. In addition, throughout the specification,the meaning of “a”, “an”, and “the” include plural references. Themeaning of “in” includes “in” and “on.”

As used herein the term, “in vitro” refers to an artificial environmentand to processes or reactions that occur within an artificialenvironment. In vitro environments may include, but are not limited to,test tubes and cell cultures. The term “in vivo” refers to the naturalenvironment (e.g., an animal or a cell) and to processes or reactionsthat occur within a natural environment.

As used herein, the terms “subject” and “patient” refer to any animal,such as a mammal like a dog, cat, bird, livestock, and preferably ahuman (e.g., a human with a disease such as obesity, diabetes, orinsulin resistance). Besides being useful for human treatment, certaincompounds and formulations disclosed herein may also be useful forveterinary treatment of companion animals, exotic animals and farmanimals, including mammals, rodents, and the like. More preferredanimals include horses, dogs, and cats.

As used herein, the term “effective amount” refers to the amount of acomposition sufficient to effect beneficial or desired results. Aneffective amount can be administered in one or more administrations,applications, or dosages and is not intended to be limited to aparticular formulation or administration route.

As used herein, the term “administration” refers to the act of giving adrug, prodrug, or other agent, or therapeutic treatment to a subject.Exemplary routes of administration to the human body can be through theeyes (ophthalmic), mouth (oral), skin (transdermal, topical), nose(nasal), lungs (inhalant), oral mucosa (buccal), ear, by injection(e.g., intravenously, subcutaneously, intratumorally, intraperitoneally,etc.), and the like.

As used herein, the term “co-administration” refers to theadministration of at least two agents or therapies to a subject. In someembodiments, the co-administration of two or more agents or therapies isconcurrent. In other embodiments, a first agent/therapy is administeredprior to a second agent/therapy. Those of skill in the art understandthat the formulations and/or routes of administration of the variousagents or therapies used may vary. The appropriate dosage forco-administration can be readily determined by one skilled in the art.In some embodiments, when agents or therapies are co-administered, therespective agents or therapies are administered at lower dosages thanappropriate for their administration alone. Thus, co-administration isespecially desirable in embodiments where the co-administration of theagents or therapies lowers the requisite dosage of a potentially harmful(e.g., toxic) agent.

As used herein, the term “pharmaceutical composition” refers to thecombination of an active agent with a carrier, inert or active, makingthe composition especially suitable for therapeutic use.

The terms “pharmaceutically acceptable” or “pharmacologicallyacceptable”, as used herein, refer to compositions that do notsubstantially produce adverse reactions, e.g., toxic, allergic, orimmunological reactions, when administered to a subject.

As used herein, the term “treating” includes reducing or alleviating atleast one adverse effect or symptom of a disease or disorder throughintroducing in any way a therapeutic composition of the presenttechnology into or onto the body of a subject. “Treatment” refers toboth therapeutic treatment and prophylactic or preventative measures,wherein the object is to prevent or slow down (lessen) the targetedpathologic condition or disorder. Those in need of treatment includethose already with the disorder as well as those prone to have thedisorder or those in whom the disorder is to be prevented.

As used herein, “therapeutically effective dose” refers to an amount ofa therapeutic agent sufficient to bring about a beneficial or desiredclinical effect. Said dose can be administered in one or moreadministrations. However, the precise determination of what would beconsidered an effective dose may be based on factors individual to eachpatient, including, but not limited to, the patient's age, size, type orextent of disease, stage of the disease, route of administration, thetype or extent of supplemental therapy used, ongoing disease process,and type of treatment desired (e.g., aggressive versus conventionaltreatment).

As used herein, the term “sample” is used in its broadest sense. In onesense, it is meant to include a specimen or culture obtained from anysource, as well as biological and environmental samples. Biologicalsamples may be obtained from animals (including humans) and encompassfluids, solids, tissues, and gases. Biological samples include bloodproducts, such as plasma, serum and the like. Environmental samplesinclude environmental material such as surface matter, soil, water, andindustrial samples. Such examples are not however to be construed aslimiting the sample types applicable to the present technology.

The term “deuterium enrichment” refers to the percentage ofincorporation of deuterium at a given position in a molecule in theplace of hydrogen. For example, deuterium enrichment of 1% at a givenposition means that 1% of molecules in a given sample contain deuteriumat the specified position. Because the naturally occurring distributionof deuterium is about 0.0156%, deuterium enrichment at any position incompound synthesized using non-enriched starting materials is about0.0156%. The deuterium enrichment can be determined using conventionalanalytical methods known to one of ordinary skill in the art, includingmass spectrometry and nuclear magnetic resonance spectroscopy.

The term “is/are deuterium”, when used to describe a given position in amolecule or the symbol “D”, when used to represent a given position in adrawing of a molecular structure, means that the specified position isenriched with deuterium above the naturally occurring distribution ofdeuterium. In one embodiment deuterium enrichment is no less than about1%, in another no less than about 5%, in another no less than about 10%,in another no less than about 20%, in another no less than about 50%, inanother no less than about 70%, in another no less than about 80%, inanother no less than about 90%, or in another no less than about 98% ofdeuterium at the specified position.

As used herein, the terms “alkyl” and the prefix “alk-” are inclusive ofboth straight chain and branched chain saturated or unsaturated groups,and of cyclic groups, e.g., cycloalkyl and cycloalkenyl groups. Unlessotherwise specified, acyclic alkyl groups are from 1 to 6 carbons.Cyclic groups can be monocyclic or polycyclic and preferably have from 3to 8 ring carbon atoms. Exemplary cyclic groups include cyclopropyl,cyclopentyl, cyclohexyl, and adamantyl groups. Alkyl groups may besubstituted with one or more substituents or unsubstituted. Exemplarysubstituents include alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio,halogen, alkylsilyl, hydroxyl, fluoroalkyl, perfluoralkyl, amino,aminoalkyl, disubstituted amino, quaternary amino, hydroxyalkyl,carboxyalkyl, and carboxyl groups. When the prefix “alk” is used, thenumber of carbons contained in the alkyl chain is given by the rangethat directly precedes this term, with the number of carbons containedin the remainder of the group that includes this prefix definedelsewhere herein. For example, the term “C₁-C₄ alkaryl” exemplifies anaryl group of from 6 to 18 carbons (e.g., see below) attached to analkyl group of from 1 to 4 carbons.

As used herein, the term “aryl” refers to a carbocyclic aromatic ring orring system. Unless otherwise specified, aryl groups are from 6 to 18carbons. Examples of aryl groups include phenyl, naphthyl, biphenyl,fluorenyl, and indenyl groups.

As used herein, the term “heteroaryl” refers to an aromatic ring or ringsystem that contains at least one ring heteroatom (e.g., O, S, Se, N, orP). Unless otherwise specified, heteroaryl groups are from 1 to 9carbons. Heteroaryl groups include furanyl, thienyl, pyrrolyl,imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl,triazolyl, tetrazolyl, oxadiazolyl, oxatriazolyl, pyridyl, pyridazyl,pyrimidyl, pyrazyl, triazyl, benzofuranyl, isobenzofuranyl,benzothienyl, indole, indazolyl, indolizinyl, benzisoxazolyl,quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, naphtyridinyl,phthalazinyl, phenanthrolinyl, purinyl, and carbazolyl groups.

As used herein, the term “heterocycle” refers to a non-aromatic ring orring system that contains at least one ring heteroatom (e.g., O, S, Se,N, or P). Unless otherwise specified, heterocyclic groups are from 2 to9 carbons. Heterocyclic groups include, for example, dihydropyrrolyl,tetrahydropyrrolyl, piperazinyl, pyranyl, dihydropyranyl,tetrahydropyranyl, dihydrofuranyl, tetrahydrofuranyl, dihydrothiophene,tetrahydrothiophene, and morpholinyl groups.

Aryl, heteroaryl, or heterocyclic groups may be unsubstituted orsubstituted by one or more substituents selected from the groupconsisting of C₁₋₆ alkyl, hydroxy, halo, nitro, C₁₋₆ alkoxy, C₁₋₆alkylthio, trifluoromethyl, C₁₋₆ acyl, arylcarbonyl, heteroarylcarbonyl,nitrile, C₁₋₆ alkoxycarbonyl, alkaryl (where the alkyl group has from 1to 4 carbon atoms), and alkheteroaryl (where the alkyl group has from 1to 4 carbon atoms).

As used herein, the term “alkoxy” refers to a chemical substituent ofthe formula —OR, where R is an alkyl group. By “aryloxy” is meant achemical substituent of the formula —OR′, where R′ is an aryl group.

As used herein, the term “C_(x-y) alkaryl” refers to a chemicalsubstituent of formula —RR′, where R is an alkyl group of x to y carbonsand R′ is an aryl group as defined elsewhere herein.

As used herein, the term “C_(x-y) alkheteraryl” refers to a chemicalsubstituent of formula RR″, where R is an alkyl group of x to y carbonsand R″ is a heteroaryl group as defined elsewhere herein.

As used herein, the term “halide” or “halogen” or “halo” refers tobromine, chlorine, iodine, or fluorine.

As used herein, the term “non-vicinal O, S, or N” refers to an oxygen,sulfur, or nitrogen heteroatom substituent in a linkage, where theheteroatom substituent does not form a bond to a saturated carbon thatis bonded to another heteroatom.

The compounds disclosed herein can exist as therapeutically acceptablesalts. The term ‘pharmaceutically acceptable salt”, as used herein,represents salts or zwitterionic forms of the compounds disclosed hereinwhich are therapeutically acceptable as defined herein. The salts can beprepared during the final isolation and purification of the compounds orseparately by reacting the appropriate compound with a suitable acid orbase. Therapeutically acceptable salts include acid and basic additionsalts. For a more complete discussion of the preparation and selectionof salts, refer to Handbook of Pharmaceutical Salts, Properties, andUse, Stah and Wermuth, Ed., (Wiley-VCH and VHCA, Zurich, 2002) and Bergeet al. (1977) J Pharm. Sci. 66: 1-19.

Suitable acids for use in the preparation of pharmaceutically acceptablesalts include, but are not limited to, acetic acid, 2,2-dichloroaceticacid, acylated amino acids, adipic acid, alginic acid, ascorbic acid,L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoicacid, boric acid, (+)-camphoric acid, camphorsulfonic acid,(+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylicacid, cinnamic acid, citric acid, cyclamic acid, cyclohexanesulfamicacid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonicacid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid,galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid,D-glucuronic acid, L-glutamic acid, alpha-oxo-glutaric acid, glycolicacid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodicacid, (+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, lauricacid, maleic acid, (−)-L-malic acid, malonic acid, (±)-DL-mandelic acid,methanesulfonic acid, naphthalene-2-sulfonic acid,naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinicacid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid,pamoic acid, perchloric acid, phosphoric acid, L-pyroglutamic acid,saccharic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid,stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaricacid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, andvaleric acid.

Suitable bases for use in the preparation of pharmaceutically acceptablesalts, including, but not limited to, inorganic bases, such as magnesiumhydroxide, calcium hydroxide, potassium hydroxide, zinc hydroxide, orsodium hydroxide; and organic bases, such as primary, secondary,tertiary, and quaternary, aliphatic and aromatic amines, includingL-arginine, benethamine, benzathine, choline, deanol, diethanolamine,diethylamine, dimethylamine, dipropylamine, diisopropylamine,2-(diethylamino)-ethanol, ethanolamine, ethylamine, ethylenediamine,isopropylamine, N-methyl-glucamine, hydrabamine, 1H-imidazole, L-lysine,morpholine, 4-(2-hydroxyethyl)-morpholine, methylamine, piperidine,piperazine, propylamine, pyrrolidine, 1-(2-hydroxyethyl)-pyrrolidine,pyridine, quinuclidine, quinoline, isoquinoline, secondary amines,triethanolamine, trimethylamine, triethylamine, N-methyl-D-glucamine,2-amino-2-(hydroxymethyl)-1,3-propanediol, and tromethamine.

For structural representations where the chirality of a carbon has beenleft unspecified it is to be presumed by one skilled in the art thateither chiral form of that stereocenter is possible.

EMBODIMENTS OF THE TECHNOLOGY

Deuterium Kinetic Isotope Effect

To eliminate foreign substances such as therapeutic agents, the animalbody expresses various enzymes, such as the cytochrome P450 enzymes(CYPs), esterases, proteases, reductases, dehydrogenases, and monoamineoxidases, to react with and convert these foreign substances to morepolar intermediates or metabolites for renal excretion. Such metabolicreactions frequently involve the oxidation of a carbon-hydrogen (C—H)bond to either a carbon-oxygen (C—O) or a carbon-carbon (C—C) π-bond.The resultant metabolites may be stable or unstable under physiologicalconditions, and can have substantially different pharmacokinetic,pharmacodynamic, and acute and long-term toxicity profiles relative tothe parent compounds. For most drugs, such oxidations are generallyrapid and ultimately lead to administration of multiple or high dailydoses.

The relationship between the activation energy and the rate of reactionmay be quantified by the Arrhenius equation given by k=Ae^(−E) ^(act)^(/(RT)), which describes the dependence of the rate constant k for thechemical reaction on the absolute temperature T (in Kelvin), where A isthe “pre-exponential factor”, E_(act) is the activation energy, and R isthe universal gas constant. The Arrhenius equation states that, at agiven temperature, the rate of a chemical reaction depends exponentiallyon the activation energy (E_(act)).

The transition state in a reaction is a short lived state along thereaction pathway during which the original bonds have stretched to theirlimit. By definition, the activation energy E_(act) for a reaction isthe energy required to reach the transition state of that reaction. Oncethe transition state is reached, the molecules can either revert to theoriginal reactants or form new bonds giving rise to reaction products. Acatalyst facilitates a reaction process by lowering the activationenergy leading to a transition state. Enzymes are examples of biologicalcatalysts.

Carbon-hydrogen bond strength is directly proportional to the absolutevalue of the ground-state vibrational energy of the bond. Thisvibrational energy depends on the mass of the atoms that form the bondand increases as the mass of one or both of the atoms making the bondincreases. Since deuterium (D) has twice the mass of “regular” hydrogen,also called protium (¹H), a C-D bond is approximately 10 times strongerthan the corresponding C—¹H bond, making it more resistant to chemicalor enzymatic cleavage. Consequently, if a C—¹H bond is broken during arate-determining step in a chemical reaction (e.g., the step with thehighest transition state energy), then substituting a deuterium for thatprotium will cause a decrease in the reaction rate. This phenomenon isknown as the deuterium kinetic isotope effect (DKIE). The magnitude ofthe DKIE can be expressed as the ratio between the rates of a givenreaction in which a C—¹H bond is broken and the same reaction wheredeuterium is substituted for protium. The DKIE can range from about 1(no isotope effect) to very large numbers such as 50 or more.Substitution of tritium for hydrogen results in yet a stronger bond thandeuterium and gives numerically larger isotope effects.

Deuterium (²H or D) is a stable and non-radioactive isotope of hydrogenthat has approximately twice the mass of protium (¹H), the most commonisotope of hydrogen. Deuterium oxide (D₂O or “heavy water”) looks andtastes like H₂O, but it has different physical properties.

Deuteration of pharmaceuticals to improve pharmacokinetics (PK),pharmacodynamics (PD), and toxicity profiles has been demonstratedpreviously with some classes of drugs. For example, the DKIE was used todecrease the hepatotoxicity of halothane, presumably by limiting theproduction of reactive species such as trifluoroacetyl chloride.However, this method is not applicable to all drug classes and thus thebiological activity of a deuterated drug in vivo is not predictable. Forexample, deuterium incorporation can lead to metabolic switching anddeuterium-protium exchange in vivo. Metabolic switching occurs whenmetabolism is hindered at one site and the resulting suppression of onemetabolic pathway promotes metabolism at another site. Accordingly,metabolic switching can lead to different proportions of knownmetabolites as well as altogether new metabolites. This new metabolicprofile may impart more or less toxicity. Deuterium-protium exchangewithin the physiological environment leads to loss of the deuteratedform and thus minimizes the advantages of deuteration in vivo. Suchproblems are non-obvious and are not predictable a priori for any drugclass.

In some embodiments deuterated amlexanox derivatives were synthesizedand their biological activity tested. In particular, amlexanox ismetabolized in humans by oxidation at the isopropyl group to produce thefollowing compound (see, e.g., Kuriki et al. (1985) “Antiallergic actionof amoxanox (AA-673), its main metabolite M-I and tranilast” Yakuri toChiryo (1973-2000) 13(11): 6435-46):

The present technology was developed to inhibit metabolism at this site.Producing an amlexanox medicine with a longer half-life was contemplatedto produce a compound having a greater efficacy and cost savings. Inparticular, the compounds provided herein are contemplated to reduce oreliminate unwanted metabolites, increase the half-life of the drug,decrease the number of doses needed to achieve a desired effect,decrease the amount of a dose needed to achieve a desired effect,increase the formation of active metabolites, decrease the production ofdeleterious metabolites in specific tissues, and/or create a moreeffective drug and/or a safer drug.

In some embodiments, certain compounds disclosed herein possess usefulkinase inhibiting activity and may be used in the treatment orprophylaxis of a disorder in which a kinase plays an active role. Thus,certain embodiments also provide pharmaceutical compositions comprisingone or more compounds disclosed herein together with a pharmaceuticallyacceptable carrier, as well as methods of making and using the compoundsand compositions. Some embodiments provide methods for inhibiting akinase activity. Other embodiments provide methods for treating akinase-mediated disorder in a patient in need of such treatment, e.g.,comprising administering to said patient a therapeutically effectiveamount of a compound or composition according to the present invention.Also provided is the use of certain compounds disclosed herein for usein the manufacture of a medicament for the prevention or treatment of adisorder ameliorated by inhibiting a kinase activity.

The compounds as disclosed herein may also contain less prevalentisotopes for other elements, including, but not limited to, ¹³C or ¹⁴Cfor carbon, ³³S, ³⁴S, or ³⁶S for sulfur, ¹⁵N for nitrogen, and ¹⁷O or¹⁸O for oxygen.

In some embodiments, the compound disclosed herein may expose a patientto D₂O or DHO, e.g., about 0.000005% D₂O or about 0.00001% DHO, assumingthat all of the C-D bonds in the compounds as disclosed herein aremetabolized and released as D₂O or DHO. In certain embodiments, thelevels of D₂O shown to cause toxicity in animals is much greater thaneven the maximum limit of exposure caused by administration of thedeuterium compound as disclosed herein. Thus, in certain embodiments,the deuterium-compounds disclosed herein do not cause any additionaltoxicity due to the formation of D₂O or DHO upon drug metabolism.

The deuterated compounds disclosed herein maintain the beneficialaspects of the corresponding non-isotopically enriched amlexanoxmolecules while substantially increasing the maximum tolerated dose,decreasing toxicity, increasing the half-life (T_(1/2)), lowering themaximum plasma concentration (C_(max)) of the minimum efficacious dose(MED), lowering the efficacious dose and thus decreasing thenon-mechanism-related toxicity, and/or lowering the probability ofdrug-drug interactions.

Deuterated Amlexanox

The carbon-hydrogen bonds of amlexanox contain a naturally occurringdistribution of hydrogen isotopes, namely ¹H or protium (about99.9844%), ²H or deuterium (about 0.0156%), and ³H or tritium (in therange between about 0.5 and 67 tritium atoms per 10¹⁸ protium atoms).The compounds provided herein have increased levels of deuterium atrelevant sites of oxidation to thus produce a deuterium kinetic isotopeeffect that improves the pharmacokinetic, pharmacologic, and/ortoxicologic profiles of amlexanox in comparison with amlexanox havingnaturally occurring levels of deuterium.

Accordingly, some embodiments provide a deuterated amlexanox, aderivative thereof, or a pharmaceutically acceptable salt thereof.Conventional amlexanox and its synthesis are described in U.S. Pat. No.4,143,042, herein incorporated by reference in its entirety. Amlexanox(2-amino-7-isopropyl-1-azaxanthone-3-carboxylic acid;2-amino-7-isopropyl-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxylic acid)has a CAS Number of 68302-57-8, a molecular weight of 298.3, and issynthesized as a crystalline solid.

However, no obvious and/or direct synthetic route is available toprovide deuterated amlexanox using conventional amlexanox as a startingmaterial. Thus, the technology described herein provides deuteratedamlexanox and methods for producing deuterated amlexanox de novo usingnovel synthetic schemes to construct amlexanox from smaller deuteratedcompounds (see Examples).

In some embodiments, the compound has the structure of Formula I:

in which X, Y, or Z is a group that comprises a deuterium (D).

The compound of general Formula (I) can be converted to thecorresponding organic amine salts, alkali metal salts, or ammonium saltsby reacting (I) in the per se conventional manner with an organic amine(e.g., ethanolamine, diethanolamine, dl-methylephedrin,1-(3,5-dihydroxyphenyl)-L-isopropylaminoethanol, isoproterenol,dextromethorphan, hetrazan (diethylcarbamazine), diethylamine,triethylamine, etc.), an alkali metal hydroxide (e.g., sodium hydroxide,potassium hydroxide, etc.) or ammonia, for example by mixing themtogether and heating in a suitable solvent.

Pharmacological Compositions

While it may be possible for the compounds of the subject invention tobe administered as the raw chemical, it is also possible to present themas a pharmaceutical composition. Accordingly, provided herein arepharmaceutical compositions which comprise one or more of certaincompounds disclosed herein, or one or more pharmaceutically acceptablesalts, prodrugs, or solvates thereof, together with one or morepharmaceutically acceptable carriers thereof and optionally one or moreother therapeutic ingredients. It is generally contemplated that thecompounds according to the technology provided are formulated foradministration to a mammal, and especially to a human with a conditionthat is responsive to the administration of such compounds. Therefore,where contemplated compounds are administered in a pharmacologicalcomposition, it is contemplated that the contemplated compounds areformulated in admixture with a pharmaceutically acceptable carrier. Forexample, contemplated compounds can be administered orally aspharmacologically acceptable salts, or intravenously in a physiologicalsaline solution (e.g., buffered to a pH of about 7.2 to 7.5).Conventional buffers such as phosphates, bicarbonates, or citrates canbe used for this purpose. Of course, one of ordinary skill in the artmay modify the formulations within the teachings of the specification toprovide numerous formulations for a particular route of administration.In particular, contemplated compounds may be modified to render themmore soluble in water or other vehicle, which for example, may be easilyaccomplished with minor modifications (e.g., salt formulation,esterification, etc.) that are well within the ordinary skill in theart. It is also well within the ordinary skill of the art to modify theroute of administration and dosage regimen of a particular compound tomanage the pharmacokinetics of the present compounds for maximumbeneficial effect in a patient.

With respect to administration to a subject, it is contemplated that thecompounds be administered in a pharmaceutically effective amount. One ofordinary skill recognizes that a pharmaceutically effective amountvaries depending on the therapeutic agent used, the subject's age,condition, and sex, and on the extent of the disease in the subject.Generally, the dosage should not be so large as to cause adverse sideeffects, such as hyperviscosity syndromes, pulmonary edema, congestiveheart failure, and the like. The dosage can also be adjusted by theindividual physician or veterinarian to achieve the desired therapeuticgoal.

As used herein, the actual amount encompassed by the term“pharmaceutically effective amount” will depend on the route ofadministration, the type of subject being treated, and the physicalcharacteristics of the specific subject under consideration. Thesefactors and their relationship to determining this amount are well knownto skilled practitioners in the medical, veterinary, and other relatedarts. This amount and the method of administration can be tailored toachieve optimal efficacy but will depend on such factors as weight,diet, concurrent medication, and other factors that those skilled in theart will recognize.

In some embodiments, deuterated amlexanox, a derivative thereof, or apharmaceutically acceptable salt thereof, is administered in apharmaceutically effective amount. In some embodiments, deuteratedamlexanox, a derivative thereof, or a pharmaceutically acceptable saltthereof, is administered in a therapeutically effective dose.

The dosage amount and frequency are selected to create an effectivelevel of the compound without substantially harmful effects. Whenadministered orally or intravenously, the dosage of deuterated amlexanoxor related compounds will generally range from 0.001 to 10,000 mg/kg/dayor dose (e.g., 0.01 to 1000 mg/kg/day or dose; 0.1 to 100 mg/kg/day ordose). In some exemplary embodiments, the deuterated amlexanox isadministered in 1 or more 2-mg doses. In some exemplary embodiments, thedeuterated amlexanox is administered as a 5% paste.

Methods of administering a pharmaceutically effective amount include,without limitation, administration in parenteral, oral, intraperitoneal,intranasal, topical, sublingual, rectal, and vaginal forms. Parenteralroutes of administration include, for example, subcutaneous,intravenous, intramuscular, intrastemal injection, and infusion routes.In some embodiments, amlexanox, a derivative thereof, or apharmaceutically acceptable salt thereof, is administered orally.

Pharmaceutical compositions preferably comprise one or more compounds ofthe present technology associated with one or more pharmaceuticallyacceptable carriers, diluents, or excipients. Pharmaceuticallyacceptable carriers are known in the art such as those described in, forexample, Remingtons Pharmaceutical Sciences, Mack Publishing Co. (A. R.Gennaro edit. 1985), explicitly incorporated herein by reference for allpurposes.

Amlexanox has been used as an oral tablet (e.g., 25-mg tablets) in Japanfor treatment of bronchial asthma and as a topical oral paste in theUnited States (Aphthasol) for treatment of aphthous ulcers (cankersores). In some embodiments, deuterated amlexanox prepared similarly toeither of these formulations may be used for the indications describedherein. In other embodiments, different formulations are used. Aphthasolcontains 5% amlexanox in an adhesive oral paste. Each gram of beigecolored oral paste contains 50 mg of amlexanox in an adhesive oral pastebase consisting of benzyl alcohol, gelatin, glyceryl monostearate,mineral oil, pectin, petrolatum, and sodium carboxymethylcellulose.Accordingly, the technology contemplates deuterated amlexanox providedin similar formulations.

In some embodiments, a single dose of deuterated amlexanox or a relatedcompound is administered to a subject. In other embodiments, multipledoses are administered over two or more time points, separated by hours,days, weeks, etc. In some embodiments, compounds are administered over along period of time (e.g., chronically), for example, for a period ofmonths or years (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or moremonths or years). In such embodiments, compounds may be taken on aregular scheduled basis (e.g., daily, weekly, etc.) for the duration ofthe extended period.

The present technology generally relates to therapeutic compositions andformulations comprising deuterated amlexanox. More particularly, thepresent technology relates to an oral medicament, a dietary supplement,a nutritional supplement, a food supplement, a food additive, apharmaceutical, a nutraceutical, or nutratherapeutical formulation.

Combination Therapy

The compounds disclosed herein may also be combined or used incombination with other agents useful in the treatment of kinase-mediateddisorders. Or, by way of example only, the therapeutic effectiveness ofone of the compounds described herein may be enhanced by administrationof an adjuvant (e.g., by itself the adjuvant may only have minimaltherapeutic benefit, but in combination with another therapeutic agent,the overall therapeutic benefit to the patient is enhanced).

Such other agents, adjuvants, or drugs, may be administered, by a routeand in an amount commonly used therefor, simultaneously or sequentiallywith a compound as disclosed herein. When a compound as disclosed hereinis used contemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compounddisclosed herein may be utilized, but is not required.

In some embodiments, a compound provided herein is combined with ananti-obesity and/or an anti-diabetes therapy. For example, in someembodiments a compound described herein is provided with a meglitinide,e.g., to stimulate the release of insulin. Exemplary meglitinides arerepaglinide (Prandin) and nateglinide (Starlix). In some embodiments, acompound described herein is provided with a sulfonylurea, e.g., tostimulate the release of insulin. Exemplary sulfonylureas are glipizide(Glucotrol), glimepiride (Amaryl), and glyburide (DiaBeta, Glynase). Insome embodiments, a compound described herein is provided with adipeptidyl peptidase-4 (DPP-4) inhibitor, e.g., to stimulate the releaseof insulin and/or to inhibit the release of glucose from the liver.Exemplary dipeptidyl peptidase-4 (DPP-4) inhibitors are saxagliptin(Onglyza), sitagliptin (Januvia), and linagliptin (Tradjenta). In someembodiments, a compound described herein is provided with a biguanide,e.g., to inhibit the release of glucose from the liver and/or to improvesensitivity to insulin. An exemplary biguanide is metformin (Fortamet,Glucophage). In some embodiments, a compound described herein isprovided with a thiazolidinedione, e.g., to improve sensitivity toinsulin and/or to inhibit the release of glucose from the liver.Exemplary thiazolidinediones include but are not limited torosiglitazone (Avandia) and pioglitazone (Actos). In some embodiments acompound described herein is provided with an alpha-glucosidaseinhibitor, e.g., to slow the breakdown of starches and some sugars.Exemplary alpha-glucosidase inhibitors include acarbose (Precose) andmiglitol (Glyset). In some embodiments, a compound as described hereinis provided with an injectable medication such as an amylin mimetic oran incretin memetic, e.g., to stimulate the release of insulin. Anexemplary amylin mimetic is pramlintide (Symlin); exemplary incretinmimetics include exenatide (Byetta) and liraglutide (Victoza). In someembodiments a compound described herein is provided with insulin. Thetechnology is not limited to any particular form of insulin, butencompasses providing the compounds described with any form of insulin.In some embodiments, the compounds described are used with an insulininjection. In some embodiments, a compound described herein is providedwith more than one additional therapy (e.g., drug or other biologicallyactive composition or compound), e.g., two, three, four or morecompounds.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more non-steroidal anti-inflammatory agents, anilideanalgesics, glucocorticoids, and immunosuppressants.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more non-steroidal anti-inflammatory agents, including, butnot limited to, aceclofenac, acemetacin, amoxiprin, aspirin,azapropazone, benorilate, bromfenac, carprofen, celecoxib, cholinemagnesium salicylate, diclofenac, diflunisal, etodolac, etoracoxib,faislamine, fenbuten, fenoprofen, flurbiprofen, ibuprofen, indometacin,ketoprofen, ketorolac, lornoxicam, loxoprofen, lumiracoxib, meloxicam,meclofenamic acid, mefenamic acid, meloxicam, metamizole, methylsalicylate, magnesium salicylate, nabumetone, naproxen, nimesulide,oxyphenbutazone, parecoxib, phenylbutazone, piroxicam, salicylsalicylate, sulindac, sulfinprazone, suprofen, tenoxicam, tiaprofenicacid, and tolmetin. In certain embodiments, the compounds disclosedherein can be combined with one or more anilide analgesics, including,but not limited to, acetaminophen and phenacetin.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more glucocorticoids, including, but not limited to,beclometasone, budesonide, flunisolide, betamethasone, fluticasone,triamcinolone, mometasone, ciclesonide, hydrocortisone, cortisoneacetate, prednisone, prednisolone, methylprednisolone, anddexamethasone.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more immunosuppressants, including, but not limited to,fingolimod, cyclosporine A, azathioprine, dexamethasone, tacrolimus,sirolimus, pimecrolimus, mycophenolate salts, everolimus, basiliximab,daclizumab, anti-thymocyte globulin, anti-lymphocyte globulin, andCTLA4IgG.

In some embodiments, deuterated amlexanox, a derivative thereof, or apharmaceutically acceptable salt thereof, is co-administered with one ormore additional therapeutic agents or medical interventions. In someembodiments, co-administration involves co-formulation of two or moreagents together into the same medicament. In other embodiments, theagents are in separate formulations but are administered together,either simultaneously or in sequence (e.g., separated by one or moreminutes, hours, days, etc.). In some embodiments, where a synergistic oradditive benefit is achieved, the co-administered agent may be providedat a lower dose than would normally be administered if that agent werebeing used in isolation to treat the disease or condition.

The compounds disclosed herein can also be administered in combinationwith other classes of compounds, including, but not limited to,norepinephrine reuptake inhibitors (NRIs) such as atomoxetine; dopaminereuptake inhibitors (DARIs), such as methylphenidate;serotonin-norepinephrine reuptake inhibitors (SNRIs), such asmilnacipran; sedatives, such as diazepham; norepinephrine-dopaminereuptake inhibitor (NDRIs), such as bupropion;serotonin-norepinephrine-dopamine-reuptake-inhibitors (SNDRIs), such asvenlafaxine; monoamine oxidase inhibitors, such as selegiline;hypothalamic phospholipids; endothelin converting enzyme (ECE)inhibitors, such as phosphoramidon; opioids, such as tramadol;thromboxane receptor antagonists, such as ifetroban; potassium channelopeners; thrombin inhibitors, such as hirudin; hypothalamicphospholipids; growth factor inhibitors, such as modulators of PDGFactivity; platelet activating factor (PAF) antagonists; anti-plateletagents, such as GPIIb/IIIa blockers (e.g., abdximab, eptifibatide, andtirofib an), P2Y(AC) antagonists (e.g., clopidogrel, ticlopidine andCS-747), and aspirin; anticoagulants, such as warfarin; low molecularweight heparins, such as enoxaparin; Factor VIIa Inhibitors and FactorXa Inhibitors; renin inhibitors; neutral endopeptidase (NEP) inhibitors;vasopepsidase inhibitors (dual NEP-ACE inhibitors), such as omapatrilatand gemopatrilat; HMG CoA reductase inhibitors, such as pravastatin,lovastatin, atorvastatin, simvastatin, NK-104 (a.k.a. itavastatin,nisvastatin, or nisbastatin), and ZD-4522 (also known as rosuvastatin,or atavastatin or visastatin); squalene synthetase inhibitors; fibrates;bile acid sequestrants, such as questran; niacin; anti-atheroscleroticagents, such as ACAT inhibitors; MTP Inhibitors; calcium channelblockers, such as amlodipine besylate; potassium channel activators;alpha-muscarinic agents; beta-muscarinic agents, such as carvedilol andmetoprolol; antiarrhythmic agents; diuretics, such as chlorothlazide,hydrochlorothiazide, flumethiazide, hydroflumethiazide,bendroflumethiazide, methylchlorothiazide, trichloromethiazide,polythiazide, benzothlazide, ethacrynic acid, tricrynafen,chlorthalidone, furosenilde, musolimine, bumetanide, triamterene,amiloride, and spironolactone; thrombolytic agents, such as tissueplasminogen activator (tPA), recombinant tPA, streptokinase, urokinase,prourokinase, and anisoylated plasminogen streptokinase activatorcomplex (APSAC); anti-diabetic agents, such as biguanides (e.g.metformin), glucosidase inhibitors (e.g., acarbose), insulins,meglitinides (e.g., rep aglinide), sulfonylureas (e.g., glimepiride,glyburide, and glipizide), thiozolidinediones (e.g. troglitazone,rosiglitazone and pioglitazone), and PPAR-gamma agonists;mineralocorticoid receptor antagonists, such as spironolactone andeplerenone; growth hormone secretagogues; aP2 inhibitors;phosphodiesterase inhibitors, such as PDE III inhibitors (e.g.,cilostazol) and PDE V inhibitors (e.g., sildenafil, tadalafil,vardenafil); protein tyrosine kinase inhibitors; antiinflammatories;antiproliferatives, such as methotrexate, FK506 (tacrolimus, Prograf),mycophenolate mofetil; chemotherapeutic agents; anticancer agents andcytotoxic agents (e.g., alkylating agents, such as nitrogen mustards,alkyl sulfonates, nitrosoureas, ethylenimines, and triazenes);antimetabolites, such as folate antagonists, purine analogues, andpyrridine analogues; antibiotics, such as anthracyclines, bleomycins,mitomycin, dactinomycin, and plicamycin; enzymes, such asL-asparaginase; farnesyl-protein transferase inhibitors; hormonalagents, such as estrogens/antiestrogens, androgens/antiandrogens,progestins, and luteinizing hormone-releasing hormone anatagonists, andoctreotide acetate; microtubule-disruptor agents, such asecteinascidins; microtubule-stabilizing agents, such as pacitaxel,docetaxel, and epothilones A-F; plant-derived products, such as vincaalkaloids, epipodophyllotoxins, and taxanes; topoisomerase inhibitors;prenyl-protein transferase inhibitors; cyclosporine; steroids, such asprednisone and dexamethasone; cytotoxic drugs, such as azathiprine andcyclophosphamide; TNF-alpha inhibitors, such as tenidap; anti-TNFantibodies or soluble TNF receptor, such as etanercept, rapamycin, andleflunimide; and cyclooxygenase-2 (COX-2) inhibitors, such as celecoxiband rofecoxib; and miscellaneous agents such as, hydroxyurea,procarbazine, mitotane, hexamethylmelamine, gold compounds, platinumcoordination complexes, such as cisplatin, satraplatin, and carboplatin.

Thus, some embodiments provide methods for treating kinase-mediateddisorders in a human or animal subject in need of such treatmentcomprising administering to said subject an amount of a compounddisclosed herein effective to reduce or prevent said disorder in thesubject, in combination with at least one additional agent for thetreatment of said disorder that is known in the art. Some embodimentsprovide therapeutic compositions comprising at least one compounddisclosed herein in combination with one or more additional agents forthe treatment of kinase-mediated disorders.

Kits

The technology provided herein also includes kits for use in the instantmethods. Kits of the technology comprise one or more containerscomprising deuterated amlexanox, a derivative thereof, or apharmaceutically acceptable salt thereof, and/or a second agent, and insome variations further comprise instructions for use in accordance withany of the methods provided herein. The kit may further comprise adescription of selecting an individual suitable treatment. Instructionssupplied in the kits of the technology are typically writteninstructions on a label or package insert (e.g., a paper insert includedwith the kit), but machine-readable instructions (e.g., instructionscarried on a magnetic or optical storage disk) are also contemplated. Insome embodiments, the kit is a package containing a sealed containercomprising any one of the preparations described above, together withinstructions for use. The kit can also include a diluent containercontaining a pharmaceutically acceptable diluent. The kit can furthercomprise instructions for mixing the preparation and the diluent. Thediluent can be any pharmaceutically acceptable diluent. Well knowndiluents include 5% dextrose solution and physiological saline solution.The container can be an infusion bag, a sealed bottle, a vial, a vialwith a septum, an ampoule, an ampoule with a septum, an infusion bag, ora syringe. The containers can optionally include indicia indicating thatthe containers have been autoclaved or otherwise subjected tosterilization techniques. The kit can include instructions foradministering the various solutions contained in the containers tosubjects.

Methods of Treatment

The technology also relates to methods of treatment with deuteratedamlexanox. According to another aspect of the technology, a method isprovided for treating a subject in need of such treatment with aneffective amount of deuterated amlexanox or a salt thereof. For example,some subjects in need of compositions according to the technology havediabetes, insulin resistance, steatosis, hepatitis, obesity, allergicrhinitis, conjunctivitis, allergy, asthma, immune disorder,atherosclerosis, canker sore, ulcer (e.g., aphthous ulcer, symptoms ofBehçet's Disease, etc.), or inflammation (e.g., inflammatory boweldisease, Crohn's disease, osteoarthritis, etc.). The method involvesadministering to the subject an effective amount of deuterated amlexanoxor salt thereof in any one of the pharmaceutical preparations describedabove, detailed herein, and/or set forth in the claims. The subject canbe any subject in need of such treatment. In the foregoing description,the technology is in connection with deuterated amlexanox or saltsthereof. Such salts include, but are not limited to, bromide salts,chloride salts, iodide salts, carbonate salts, and sulfate salts. Itshould be understood, however, that deuterated amlexanox is a member ofa class of compounds and the technology is intended to embracepharmaceutical preparations, methods, and kits containing relatedderivatives within this class. Another aspect of the technology thenembraces the foregoing summary but read in each aspect as if any suchderivative is substituted wherever “amlexanox” or “deuterated amlexanox”appears.

In some embodiments, provided herein are methods of treatmentcomprising: administering a pharmaceutically effective amount ofdeuterated amlexanox, a derivative thereof, or a pharmaceuticallyacceptable salt thereof, alone or in combination with another agent, toa subject having a condition in need of treatment. In some embodiments,the administration causes one or more of: a reduction in or eliminationof one or more symptoms of the condition, prevention of increasedseverity of one or more symptoms of the condition, and/or reduction,prevention, or elimination of further diseases or conditions.

In some embodiments, the methods provided comprise testing a subject fora disease or condition followed by administering deuterated amlexanox, aderivative thereof, or a pharmaceutically acceptable salt thereof, aloneor in combination with other agents. In some embodiments, methodscomprise administering to a subject deuterated amlexanox, a derivativethereof, or a pharmaceutically acceptable salt thereof, alone or incombination with other agents, followed by testing the subject for adisease or a condition. In some embodiments, methods comprise testing asubject for a disease or condition followed by administering deuteratedamlexanox, a derivative thereof, or a pharmaceutically acceptable saltthereof, alone or in combination with other agents, followed by a secondround of testing for a disease or condition (e.g., to monitor the effectof the treatment). In some embodiments, methods comprise testing asubject for a disease or condition followed by administering deuteratedamlexanox, a derivative thereof, or a pharmaceutically acceptable saltthereof, alone or in combination with other agents, followed by a secondround of testing for a disease or condition and a second administrationof deuterated amlexanox, a derivative thereof, or a pharmaceuticallyacceptable salt thereof, alone or in combination with other agents, withthis second administration being modified in dose, duration, frequency,or administration route in a manner dependent upon the results of theprior testing. In some embodiments, a subject is tested to assess thepresence, the absence, or the level of a disease, e.g., by assaying ormeasuring a biomarker, a metabolite, a physical symptom, an indication,etc., to determine the risk of or the presence of the disease andthereafter the subject is treated with deuterated amlexanox based on theoutcome of the test. In some embodiments, a patient is tested, treated,and then tested again to monitor the response to therapy. In someembodiments, cycles of testing and treatment may occur withoutlimitation to the pattern of testing and treating (e.g., test/treat,test/treat/test, test/treat/test/treat, test/treat/test/treat/test,test/treat/treat/test/treat/treat, etc.), the periodicity, or theduration of the interval between each testing and treatment phase.

In some embodiments, the technology provided comprises use of deuteratedamlexanox, a derivative thereof, or a pharmaceutically acceptable saltthereof, alone or in combination with other agents in the manufacture ofa medicament for the treatment of a condition. In some embodiments, thetechnology provides deuterated amlexanox, a derivative thereof, or apharmaceutically acceptable salt thereof, for the treatment of acondition.

Although the disclosure herein refers to certain illustratedembodiments, it is to be understood that these embodiments are presentedby way of example and not by way of limitation.

EXAMPLES Example 1—Synthesis of Deuterated Amlexanox

During the development of embodiments of the technology provided herein,experiments were performed to test synthetic schemes for producingdeuterated amlexanox.

A. C-7 Mono- and Hepta-Deuterated Congeners.

Scheme A delineates a route to target compounds A-9a and A-9b.Commercially available 2-deuterio-2-propanol (A-1a) and1,1,1,2,3,3,3-heptadeuterio-2-propanol (A-1b) are tosylated with tosylchloride and pyridine (Obach (2001) “Mechanism of cytochrome P4503A4-and 2D6-catalyzed dehydrogenation of ezlopitant as probed with isotopeeffects using five deuterated analogs” Drug Metab. Dispos. 29(12):1599-1607). The resulting tosylates A-2 are used to alkylate theGrignard reagent prepared from 1-bromo-4-methoxybenzene to provide theethers A-3 (Obach, supra). Demethylation of the methyl ethers iseffected with boron tribromide (Waibel et al. (2009) “Bibenzyl- andstilbene-core compounds with non-polar linker atom substituents asselective ligands for estrogen receptor beta” Eur. J. Med. Chem. 44(9):3412-3424) to afford phenols A-4. Fries rearrangement of phenols A-4with acetic anhydride and aluminum trichloride provides the acetylatedphenols A-5 (Nohara et al. 1974 “Antianaphylactic agents. I. Facilesynthesis of 4-oxo-4H-1-benzopyran-3-carboxaldehydes by Vilsmeierreagents” Tetrahedron 30(19): 3553-3561). Under Vilsmeier conditions,the phenols are cyclized to chromone aldehydes A-6 (Nohara 1974, supra).Conversion of the aldehydes to nitriles A-7 is effected withhydroxylamine and concentrated HCl (Nohara et al. (1977) “Studies onantianaphylactic agents. 5. Synthesis of 3-(1H-tetrazol-5-yl)chromones,a new series of antiallergic substances” J. Med. Chem. 20(1): 141-145).Cyclization to aminopyridine esters A-8 is accomplished by condensationwith ethyl cyanoacetate and piperidine (Nohara et al. (1985) “Studies onantianaphylactic agents. 7. Synthesis of antiallergic5-oxo-5H-[1]benzopyrano [2,3-b]pyridines” J. Med. Chem. 28(5): 559-568).Finally, deuterated amlexanox derivatives A-9a and A-9b are obtained byacidic hydrolysis of the ethyl esters (Nohara 1985, supra).

B. C-7 Dideutero Congener.

The synthesis of target compound B-4 is given in Scheme B. Compound B-1was generated in a manner according to Ukawa et al. (1985) “Synthesis ofthe metabolites and degradation products of2-amino-7-isopropyl-5-oxo-5H-[1]benzopyrano [2,3-b]pyridine-3-carboxylicacid (amoxanox)” Chem. Pharm. Bull. 33(10): 4432-7. Boc protection wascarried out under standard conditions to give B-2, which was thentreated with Wilkinson's catalyst and deuterium gas to provide dideuteroproduct B-3. A two-stage deprotection sequence (eg, sodium hydroxide tohydrolyze the ester and hydrochloric acid to cleave the Boc carbamate)provided the targeted D₂-amlexanox (B-4).

C. Alternate Scheme to C-7 Monodeutero Congener.

Compound C-1 is prepared in a manner similar to methyl2-((tert-butoxycarbonyl)amino)-5-oxo-7-(prop-1-en-2-yl)-5H-chromeno[2,3-b]pyridine-3-carboxylate(B-1) from methyl2-amino-7-isopropyl-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxylate(Nohara 1985, supra) and di-tert-butyl dicarbonate. The deuteriumexchange is performed in a similar manner described in the literature(Kurita et al. (2008) “Efficient and convenient heterogeneouspalladium-catalyzed regioselective deuteration at the benzylic position”Chem. Eur. J. 14(2): 664-673) to provide C-2. Deprotection is affordedby treatment of C-2 sequentially with aqueous sodium hydroxide withmethanol as a co-solvent followed by treatment of C-3 with 2M HCl indioxane to obtain C-4.

D. Experimental Procedures

Chemical names follow CAS nomenclature. Starting materials werepurchased from Fisher, Sigma-Aldrich Lancaster, Fluka, or TCI-Americaand were used without purification. All reaction solvents were purchasedfrom Fisher and used as received. Reactions were monitored by TLC usingpre-coated silica gel 60 F254 plates. Silica gel chromatography wasperformed with silica gel (220 to 240 mesh) obtained from Silicycle. NMRspectra were recorded on a Varian 400 MHz spectrometer. Chemical shiftsare reported in δ (parts per million), by reference to the hydrogenatedresidues of a deuterated solvent as an internal standard CDCl₃: δ=7.28(¹H NMR). Mass spectra were recorded on a Micromass LCT time-of-flightinstrument utilizing the electrospray ionization mode. Melting pointswere measured on a MEL-TEMP melting point apparatus and are uncorrected.The purity of the compounds was assessed via analytical rpHPLC with agradient of 90% A:B to 10% A:B over 6 minutes (solvent A, H₂O; solventB, acetonitrile; C18 column, 3.5 μm, 4.6×100 mm; 254 nm wavelengthdetection).

Methyl2-((tert-butoxycarbonyl)amino)-5-oxo-7-(prop-1-en-2-yl)-5H-chromeno[2,3-b]pyridine-3-carboxylate(B-2)

To a solution of methyl2-amino-5-oxo-7-(prop-1-en-2-yl)-5H-chromeno[2,3-b]pyridine-3-carboxylate(0.044 g, 0.14 mmol) in dry THF was added di-tert-butyl dicarbonate(0.036 ml, 0.16 mmol) followed by N,N-diisopropylethylamine (0.028 ml,0.16 mmol) and catalytic 4-dimethylaminopyridine. The resulting mixturewas stirred overnight at room temperature and then diluted with ethylacetate/ether, washed twice with saturated aqueous NaCl, and dried overMgSO₄. Purification by flash chromatography, eluting with 30% ethylacetate/hexane, gave B-2 (0.04 g, 68.7%) as a white solid. HPLC (tR=9.14minutes). NMR (400 MHz, CDCl₃) δ 9.34 (s, 1H), 8.35 (s, 1H), 7.94 (d,1H), 7.58 (d, 1H), 5.51 (s, 1H), 5.23 (s, 1H), 3.95 (s, 3H), 2.24 (s,3H), 1.41 (s, 9H).

Methyl2-((tert-butoxycarbonyl)amino)-7-(D₂)-isopropyl-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxylate(B-3)

Wilkinson's catalyst (chlorotris(triphenylphosphine)rhodium(I); 0.04 g,0.043 mmol) was dissolved in methyl ethyl ketone (10 ml) and deuteriumgas was bubbled into the solution via a balloon and needle. A balloon ofdeuterium gas was attached to the reaction vessel and the mixture wasstirred 1 hour before adding methyl2-((tert-butoxycarbonyl)amino)-5-oxo-7-(prop-1-en-2-yl)-5H-chromeno[2,3-b]pyridine-3-carboxylate(B-2; 0.04 g, 0.097 mmol) in methyl ethyl ketone (8 ml). The resultingmixture was stirred for 1 hour at room temperature, concentrated, andfiltered through a plug of silica gel and eluting with 30% ethylacetate/hexane. Concentration of the filtrate provided B-3 (0.035 g,87%). HPLC (tR=9.27 minutes). NMR (400 MHz, CDCl₃) δ 9.34 (s, 1H), 8.16(s, 1H), 7.68 (d, 1H), 7.55 (d, 1H), 3.95 (s, 3H), 1.40 (s, 9H), 1.31(s, 3H), 1.29 (s, 2H).

2-Amino-7-(D₂)-isopropyl-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxylicacid (B-4)

To methyl2-((tert-butoxycarbonyl)amino)-7-(D₂)-isopropyl-5-oxo-5H-chromeno[2,3-b]pyridine-3-carboxylate(B-3; 0.03 g, 0.07 mmol) dissolved in methanol (5 ml) was added aqueous1 M sodium hydroxide (0.14 ml, 0.14 mmol). The resulting mixture wasstirred overnight at room temperature and then concentrated. The residuewas dissolved in ethyl acetate (5 ml) and treated with excess 2-M HCl indioxane (5 ml). After stirring for 2 hours, the mixture was concentratedto a residue that was triturated in methanol. The solids were collectedand dried under vacuum at room temperature to give B-4 (0.012 g, 55.2%).HPLC (tR=6.48 minutes). NMR (400 MHz, CDCl₃) δ 8.88 (s, 1H), 8.24 (br s,1H), 7.90 (s, 1H), 7.65 (br s, 1H), 7.51 (d, 1H), 7.32 (d, 1H), 1.20 (s,3H), 1.19 (s, 2H). ESI+MS m/z 300.306 (M+H⁺).

Example 2—Biological Activity of Deuterated Amlexanox

During the development of embodiments of the technology provided herein,experiments were conducted to test in vitro the biological activity ofdeuterated amlexanox. In particular, data were collected to quantify theinhibitory activity of a deuterated amlexanox toward the protein kinasesIKKε and TBK1.

The effects of the compounds on these kinases were assayed in vitro. Invitro kinase assays were performed by incubating purified kinase (IKKεor TBK1) in kinase buffer containing 25 mM Tris (pH 7.5), 10 mM MgCl₂, 1mM DTT, and 10 μM ATP for 30 minutes at 30° C. in the presence of 0.5μCi γ-[³²P]-ATP and 1 μg myelin basic protein (MBP) per sample as asubstrate. Kinase inhibitors were added at 50 μM and serially diluted(final concentrations are indicated). Kinase reactions were stopped byadding 4× sodium dodecyl sulfate (SDS) sample buffer and boiling for 5minutes at 95° C. Supernatants were resolved by SDS-polyacrylamide gelelectrophoresis, transferred to nitrocellulose, and analyzed byautoradiography using a Typhoon 9410 phosphorimager (GE Lifesciences,Piscataway, N.J.). The bands were quantified using ImageQuant.

Amlexanox and deuterated (D)-amlexanox inhibited the activities of bothprotein kinases, with IC50 values in the range of 1 to 2 μM.Dose-response data for IKKε are shown in Table 1. The results show thatdeuterated amlexanox inhibits IKKε and TBK1 in a dose-dependent manner(FIG. 1).

TABLE 1 inhibition of IKKε Experiment 1 Experiment 2 Average D-AmlexanoxMBP- Normalized % MBP- Normalized % % concentration IKKε ³²P inhibitionIKKε ³²P inhibition inhibition 50.0 μM 1374.96 0.283097 71.69 1379.670.307902 69.21 70.45 25.0 μM 1720.79 0.354301 64.57 2017.15 0.45019654.99 59.78 12.5 μM 2292.47 0.472007 52.80 2636.59 0.588444 41.16 46.98 5.0 μM 2136.30 0.439852 56.01 2761.66 0.616358 38.36 47.19  1.0 μM3879.37 0.798741 20.13 4375.20 0.976473 2.35 11.24  0.5 μM 4420.640.910186 8.98 4289.95 0.957448 4.26 6.625

All publications and patents mentioned in the above specification areherein incorporated by reference in their entirety for all purposes.Various modifications and variations of the described compositions,methods, and uses of the technology will be apparent to those skilled inthe art without departing from the scope and spirit of the technology asdescribed. Although the technology has been described in connection withspecific exemplary embodiments, it should be understood that theinvention as claimed should not be unduly limited to such specificembodiments. Indeed, various modifications of the described modes forcarrying out the invention that are obvious to those skilled in the artare intended to be within the scope of the following claims.

We claim:
 1. A method of making a composition comprising deuteratedamlexanox, the method comprising: reacting an amlexanox precursor havinga structure according to

wherein R₂ comprises an alkyl, with an homogenous catalyst and deuteriumgas to provide a composition comprising deuterated amlexanox having astructure according to

wherein said composition is enriched with said deuterated amlexanoxabove the naturally occurring distribution of deuterium.
 2. The methodof claim 1 wherein said homogenous catalyst is Wilkinson's catalyst. 3.The method of claim 1 wherein said composition comprising deuteratedamlexanox is enriched more than 0.0200% with said deuterated amlexanox.4. The method of claim 1 further comprising hydrolyzing the alkyl estercomprising the R₂ group.
 5. The method of claim 1 wherein R₂ comprises amethyl or ethyl group.
 6. A method of making a deuterated amlexanoxmolecule, the method comprising: reacting an amlexanox precursormolecule having a structure according to

wherein R₂ comprises an alkyl, with an homogenous catalyst and deuteriumgas to provide a deuterated amlexanox molecule having a structureaccording to


7. The method of claim 6 wherein said homogenous catalyst is Wilkinson'scatalyst.
 8. The method of claim 6 further comprising hydrolyzing thealkyl ester comprising the R₂ group.
 9. The method of claim 6 furthercomprising hydrolyzing the alkyl ester comprising the R₂ group usingNaOH followed by HCl.
 10. The method of claim 6 wherein R₂ comprises amethyl or ethyl group.
 11. The method of claim 6 further comprisingpreparing a derivative or pharmaceutically acceptable salt of saiddeuterated amlexanox.